# Patent

## Title
Pepper hybrid BS02828008

## Abstract

The invention provides seed and plants of pepper hybrid BS02828008 and the parent lines thereof. The invention thus relates to the plants, seeds and tissue cultures of pepper hybrid BS02828008 and the parent lines thereof, and to methods for producing a pepper plant produced by crossing such plants with themselves or with another pepper plant, such as a plant of another genotype. The invention further relates to seeds and plants produced by such crossing. The invention further relates to parts of such plants, including the fruit and gametes of such plants.

## Background
CROSS-REFERENCE TO RELATED APPLICATIONS
This application claims the priority of U.S. Provisional Appl. Ser. No. 61/182,573, filed May 29, 2009, the entire disclosure of which is incorporated herein by reference.


FIELD OF THE INVENTION
The present invention relates to the field of plant breeding and, more specifically, to the development of pepper hybrid BS02828008 and the inbred pepper lines SMO28-1234 and SMO28-1254.
BACKGROUND OF THE INVENTION
The present invention relates to the field of plant breeding and, more specifically, to the development of pepper hybrid BS02828008 and the inbred pepper lines SMO28-1234 and SMO28-1254.
The goal of vegetable breeding is to combine various desirable traits in a single variety/hybrid. Such desirable traits may include any trait deemed beneficial by a grower and/or consumer, including greater yield, resistance to insects or disease, tolerance to environmental stress, and nutritional value.
Breeding techniques take advantage of a plant's method of pollination. There are two general methods of pollination: a plant self-pollinates if pollen from one flower is transferred to the same or another flower of the same plant or plant variety. A plant cross-pollinates if pollen comes to it from a flower of a different plant variety.
Plants that have been self-pollinated and selected for type over many generations become homozygous at almost all gene loci and produce a uniform population of true breeding progeny, a homozygous plant. A cross between two such homozygous plants of different genotypes produces a uniform population of hybrid plants that are heterozygous for many gene loci. Conversely, a cross of two plants each heterozygous at a number of loci produces a population of hybrid plants that differ genetically and are not uniform. The resulting non-uniformity makes performance unpredictable.
The development of uniform varieties requires the development of homozygous inbred plants, the crossing of these inbred plants, and the evaluation of the crosses. Pedigree breeding and recurrent selection are examples of breeding methods that have been used to develop inbred plants from breeding populations. Those breeding methods combine the genetic backgrounds from two or more plants or various other broad-based sources into breeding pools from which new lines and hybrids derived therefrom are developed by selfing and selection of desired phenotypes. The new lines and hybrids are evaluated to determine which of those have commercial potential.
SUMMARY OF THE INVENTION
In one aspect, the present invention provides a pepper plant of the hybrid designated BS02828008, the pepper line SMO28-1234 or pepper line SMO28-1254. Also provided are pepper plants having all the physiological and morphological characteristics of such a plant. Parts of these pepper plants are also provided, for example, including pollen, an ovule, scion, a rootstock, a fruit, and a cell of the plant.
In another aspect of the invention, a plant of pepper hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254 comprising an added heritable trait is provided. The heritable trait may comprise a genetic locus that is, for example, a dominant or recessive allele. In one embodiment of the invention, a plant of pepper hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254 is defined as comprising a single locus conversion. In specific embodiments of the invention, an added genetic locus confers one or more traits such as, for example, herbicide tolerance, insect resistance, disease resistance, and modified carbohydrate metabolism. In further embodiments, the trait may be conferred by a naturally occurring gene introduced into the genome of a line by backcrossing, a natural or induced mutation, or a transgene introduced through genetic transformation techniques into the plant or a progenitor of any previous generation thereof. When introduced through transformation, a genetic locus may comprise one or more genes integrated at a single chromosomal location.
The invention also concerns the seed of pepper hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254. The pepper seed of the invention may be provided as an essentially homogeneous population of pepper seed of pepper hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254. Essentially homogeneous populations of seed are generally free from substantial numbers of other seed. Therefore, in one embodiment of the invention, seed of hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254 may be defined as forming at least about 97% of the total seed, including at least about 98%, 99% or more of the seed. The seed population may be separately grown to provide an essentially homogeneous population of pepper plants designated BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254.
In yet another aspect of the invention, a tissue culture of regenerable cells of a pepper plant of hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254 is provided. The tissue culture will preferably be capable of regenerating pepper plants capable of expressing all of the physiological and morphological characteristics of the starting plant, and of regenerating plants having substantially the same genotype as the starting plant. Examples of some of the physiological and morphological characteristics of the hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254 include those traits set forth in the tables herein. The regenerable cells in such tissue cultures may be derived, for example, from embryos, meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistils, flowers, seed and stalks. Still further, the present invention provides pepper plants regenerated from a tissue culture of the invention, the plants having all the physiological and morphological characteristics of hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254.
In still yet another aspect of the invention, processes are provided for producing pepper seeds, plants and fruit, which processes generally comprise crossing a first parent pepper plant with a second parent pepper plant, wherein at least one of the first or second parent pepper plants is a plant of pepper line SMO28-1234 or pepper line SMO28-1254. These processes may be further exemplified as processes for preparing hybrid pepper seed or plants, wherein a first pepper plant is crossed with a second pepper plant of a different, distinct genotype to provide a hybrid that has, as one of its parents, a plant of pepper line SMO28-1234 or pepper line SMO28-1254. In these processes, crossing will result in the production of seed. The seed production occurs regardless of whether the seed is collected or not.
In one embodiment of the invention, the first step in “crossing” comprises planting seeds of a first and second parent pepper plant, often in proximity so that pollination will occur for example, mediated by insect vectors. Alternatively, pollen can be transferred manually. Where the plant is self-pollinated, pollination may occur without the need for direct human intervention other than plant cultivation.
A second step may comprise cultivating or growing the seeds of first and second parent pepper plants into plants that bear flowers. A third step may comprise preventing self-pollination of the plants, such as by emasculating the flowers (i.e., killing or removing the pollen).
A fourth step for a hybrid cross may comprise cross-pollination between the first and second parent pepper plants. Yet another step comprises harvesting the seeds from at least one of the parent pepper plants. The harvested seed can be grown to produce a pepper plant or hybrid pepper plant.
The present invention also provides the pepper seeds and plants produced by a process that comprises crossing a first parent pepper plant with a second parent pepper plant, wherein at least one of the first or second parent pepper plants is a plant of pepper hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254. In one embodiment of the invention, pepper seed and plants produced by the process are first generation (F1) hybrid pepper seed and plants produced by crossing a plant in accordance with the invention with another, distinct plant. The present invention further contemplates plant parts of such an F1 hybrid pepper plant, and methods of use thereof. Therefore, certain exemplary embodiments of the invention provide an F1 hybrid pepper plant and seed thereof.
In still yet another aspect, the present invention provides a method of producing a plant derived from hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254, the method comprising the steps of: (a) preparing a progeny plant derived from hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254, wherein said preparing comprises crossing a plant of the hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254 with a second plant; and (b) crossing the progeny plant with itself or a second plant to produce a seed of a progeny plant of a subsequent generation. In further embodiments, the method may additionally comprise: (c) growing a progeny plant of a subsequent generation from said seed of a progeny plant of a subsequent generation and crossing the progeny plant of a subsequent generation with itself or a second plant; and repeating the steps for an additional 3-10 generations to produce a plant derived from hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254. The plant derived from hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254 may be an inbred line, and the aforementioned repeated crossing steps may be defined as comprising sufficient inbreeding to produce the inbred line. In the method, it may be desirable to select particular plants resulting from step (c) for continued crossing according to steps (b) and (c). By selecting plants having one or more desirable traits, a plant derived from hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254 is obtained which possesses some of the desirable traits of the line/hybrid as well as potentially other selected traits.
In certain embodiments, the present invention provides a method of producing food or feed comprising: (a) obtaining a plant of pepper hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254, wherein the plant has been cultivated to maturity, and (b) collecting at least one pepper from the plant.
In still yet another aspect of the invention, the genetic complement of pepper hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254 is provided. The phrase “genetic complement” is used to refer to the aggregate of nucleotide sequences, the expression of which sequences defines the phenotype of, in the present case, a pepper plant, or a cell or tissue of that plant. A genetic complement thus represents the genetic makeup of a cell, tissue or plant, and a hybrid genetic complement represents the genetic make up of a hybrid cell, tissue or plant. The invention thus provides pepper plant cells that have a genetic complement in accordance with the pepper plant cells disclosed herein, and seeds and plants containing such cells.
Plant genetic complements may be assessed by genetic marker profiles, and by the expression of phenotypic traits that are characteristic of the expression of the genetic complement, e.g., isozyme typing profiles. It is understood that hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254 could be identified by any of the many well known techniques such as, for example, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang et al., 1998).
In still yet another aspect, the present invention provides hybrid genetic complements, as represented by pepper plant cells, tissues, plants, and seeds, formed by the combination of a haploid genetic complement of a pepper plant of the invention with a haploid genetic complement of a second pepper plant, preferably, another, distinct pepper plant. In another aspect, the present invention provides a pepper plant regenerated from a tissue culture that comprises a hybrid genetic complement of this invention.
In still yet another aspect, the invention provides a plant of an hybrid pepper that exhibits a combination of traits comprising a plant with a semi-spreading, semi-erect habit, with few (2-3) basal branches; an early harvest of a large number of small, somewhat angular fruit that mature from a medium-green to an orange color; a glossy fruit that typically has three locules and produces about 180 seeds per fruit, has a Brix content of about 9.7% and is suitable for fresh market sale; a fruit that averages 8.2 cm long, 5.0 cm in diameter and weighs about 73 g; a plant that appears resistant to Tobamovirus (P12) and Potato Virus Y (PVY) (P0). In certain embodiments, the combination of traits may be defined as controlled by genetic means for the expression of the combination of traits found in pepper hybrid BS02828008.
In still yet another aspect, the invention provides a method of determining the genotype of a plant of pepper hybrid BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254 comprising detecting in the genome of the plant at least a first polymorphism. The method may, in certain embodiments, comprise detecting a plurality of polymorphisms in the genome of the plant. The method may further comprise storing the results of the step of detecting the plurality of polymorphisms on a computer readable medium. The invention further provides a computer readable medium produced by such a method.
Any embodiment discussed herein with respect to one aspect of the invention applies to other aspects of the invention as well, unless specifically noted.
The term “about” is used to indicate that a value includes the standard deviation of the mean for the device or method being employed to determine the value. The use of the term “or” in the claims is used to mean “and/or” unless explicitly indicated to refer to alternatives only or the alternatives are mutually exclusive. When used in conjunction with the word “comprising” or other open language in the claims, the words “a” and “an” denote “one or more,” unless specifically noted otherwise. The terms “comprise,” “have” and “include” are open-ended linking verbs. Any forms or tenses of one or more of these verbs, such as “comprises,” “comprising,” “has,” “having,” “includes” and “including,” are also open-ended. For example, any method that “comprises,” “has” or “includes” one or more steps is not limited to possessing only those one or more steps and also covers other unlisted steps. Similarly, any plant that “comprises,” “has” or “includes” one or more traits is not limited to possessing only those one or more traits and covers other unlisted traits.
Other objects, features and advantages of the present invention will become apparent from the following detailed description. It should be understood, however, that the detailed description and any specific examples provided, while indicating specific embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.


DETAILED DESCRIPTION OF THE INVENTION
The invention provides methods and compositions relating to plants, seeds and derivatives of pepper hybrid BS02828008, pepper line SMO28-1234 and pepper line SMO28-1254. The hybrid BS02828008 is produced by the cross of parent lines SMO28-1234 and SMO28-1254. The parent lines show uniformity and stability within the limits of environmental influence. By crossing the parent lines, uniform seed of hybrid BS02828008 can be obtained.
The development of pepper hybrid BS02828008 and its parent lines can be summarized as follows.
A. Origin and Breeding History of Pepper Hybrid BS02828008
The parents of hybrid BS02828008 are SMO28-1234 and SMO28-1254. These parents were created as follows.
For parent SMO28-1234:

    Summer, Year 1 The F1 hybrid was made from the parents 2000-2029 (stake 00F-339) and 2000-2058 (stake 00F-366) and assigned the accession number 2000-6102.
    Winter, Year 2 Plants of the F1 hybrid 2000-6102 were transplanted as stake 01S-2557. The hybrid produced small, blocky, yellow fruit. Seeds of the selection, 01S-2557-3, received accession number 2001-3434.
    Summer, Year 2 Planted the F2 inbred line 2001-3434 as stake 01F-257. The hybrid developed a low vigor plant that segregated for small, blocky, yellow and orange fruit. Fruits measured about 6.8 cm long by 4.8 cm diameter, each weighed about 50 g and had a Brix of 9.0%. Seeds of the selection, 01F-257-14, tested resistant to Tobamovirus (P012), and received accession number 2001-3549.
    Winter, Year 3 Planted the F3 inbred line 2001-3549 as stake 02S-273. The line produced a small and somewhat tapered orange fruit. Fruits measured about 6.8 cm long by 4.2 cm diameter, each weighed about 40 g and had a Brix of 11.3%. Seeds of the selection, 02S-273-18, tested for resistant to the Tobamovirus (P012) and to PVY (P0), and received accession number 2002-1189.
    Summer, Year 3 Planted the F4 inbred line 2002-1189 as stake 02F-1400. The line produced small blocky and somewhat tapered orange peppers. Fruits measured about 5.8 cm long by 4.6 cm diameter, each weighed about 46 g and had a Brix of 10%. Seeds of the selection, 02F-1400-5, tested resistant to Tobamovirus (P012) and to PVY (P0), and received accession number 2002-3020.
    Winter, Year 4 Planted the F5 inbred line 2002-3020 as stake 03S-85. The line developed low vigor plants that produced small blocky and somewhat tapered orange peppers. Fruits measured about 6 cm long by 4 cm diameter, each weighed about 36 g and had a Brix of 10.6%. Seeds of the selection, 03S-85-1, tested resistant to Tobamovirus (P012) and to PVY (P0), and received accession number 2003-2079.
    Spring, Year 4 Planted the F6 inbred line 2003-2079 as stake 03U-77. The line developed low vigor but productive plants that produced small, blocky, orange peppers. Fruits measured about 6 cm long by 4.2 cm diameter, each weighed about 48 g and had a Brix of 9.6%. Seeds of the selection, 03U-77-10, tested resistant to Tobamovirus (P012) and to PVY (P0), and received accession number 2003-3276.
    Winter, Year 4 Planted the F7 inbred line 2003-3276 as stake 03F-2077. The line developed small, blocky, orange peppers. Fruits measured about 6 cm long by 4.2 cm diameter, each weighed about 39 g and had a Brix of 9.9%. Seeds of the selection, 03F-2077-M, tested resistant to Tobamovirus (P012) and to PVY (P0), and received accession number 2003-4578.
    Summer, Year 4 Accession 2003-4578 was submitted as inbred line SMO28-1234.


For parent SMO28-1254:

    Summer, Year 1 The F1 hybrid was made from the parents 2000-2033 (stake 00F-343) and 2000-2058 (stake 00F-366) and assigned the accession number 2000-6110.
    Winter, Year 2 Plants of the F1 hybrid 2000-6110 were transplanted as stake 01S-2562. The hybrid produced small, blocky, red fruit. Seeds of the selection, 01S-2562-4, received accession number 2001-3436.
    Summer, Year 2 Planted the F2 inbred line 2001-3436 as stake 01F-258. The line developed a compact plant that segregated for red and orange fruit. Fruits measured about 7.4 cm long by 5.8 cm diameter, weighed about 62 g, and had a Brix of 10.1%. Seeds of the selection, 01F-258-19, tested resistant to TMV (P012), and received accession number 2001-3556.
    Winter, Year 3 Planted the F3 inbred line 2001-3556 as stake 02S-276. The line produced small, blocky, orange fruit. Fruits measured about 7.6 cm long by 6 cm diameter, weighed about 50 g, and had a Brix of 10%. Seeds of the selection, 02S-276-5, tested resistant to TMV (P012) and to PVY (P0), and received accession number 2002-1190.
    Summer, Year 3 Planted the F4 inbred line 2002-1190 as stake 02F-1401. The line produced small, blocky, orange fruit. Fruits measured about 7.6 cm long by 5.4 cm diameter, weighed about 75 g, and had a Brix of 10.2%. Seeds of the selection, 02F-1401-6, tested resistant to TMV (P012), and received accession number 2002-3022.
    Winter, Year 4 Planted the F5 inbred line 2002-3022 as stake 03F-87. The line developed low vigor plants that produced small, blocky, orange fruit. Fruits measured about 7 cm long by 6 cm diameter, weighed about 85 g, and had a Brix of 9.6%. Seeds of the selection, 03F-87-6, tested resistant to TMV (P012), and received accession number 2003-2081.
    Summer, Year 4 Planted the F6 inbred line 2003-2081 as stake 03U-79. The line developed low vigor plants that produced small, blocky, orange fruit. Fruits measured about 7 cm long by 5.6 cm diameter, weighed about 71 g, and had a Brix of 9.3%. Seeds of the selection, 03U-79-4, tested resistant to TMV (P012) and to PVY (P0), and received accession number 2003-3278.
    Winter, Year 5 Planted the F7 inbred line 2003-3278 as stake 04S-109. The line developed low vigor plants that produced small, blocky, orange fruit. Fruits measured about 6.6 cm long by 6.6 cm diameter, weighed about 85 g, and had a Brix of 10.3%. Seeds of the selection, 04S-109-5, tested for resistant to TMV(P012) and to PVY(P0), received accession number 2004-1049.
    Summer, Year 5 Planted the F8 inbred line 2004-1049 as stake 04F-1803. The line developed compact plants that produced small, blocky, orange fruit. Fruits measured about 6.2 cm long by 5.4 cm diameter, weighed about 56 g, and had a Brix of 10.2%. Seeds of the selection, 04F-1803-M, tested resistant to TMV (P012), received accession number 2004-3047.
    Summer, Year 6 Accession 2004-3047 was submitted as inbred line SMO28-1254.


The parent lines are uniform and stable, as is a hybrid therefrom. A small percentage of variants can occur within commercially acceptable limits for almost any characteristic during the course of repeated multiplication. However no variants are expected.
B. Physiological and Morphological Characteristics of Pepper Hybrid BS02828008, Pepper Line SMO28-1234 and Pepper Line SMO28-1254
In accordance with one aspect of the present invention, there is provided a plant having the physiological and morphological characteristics of pepper hybrid BS02828008 and the parent lines thereof. A description of the physiological and morphological characteristics of such plants is presented in Tables 1-2.







TABLE 1







Physiological and Morphological Characteristics of Hybrid BS02828008













Comparison



CHARACTERISTIC
BS02828008
Variety-Boogie














1.
Species
C. annuum



2.
Maturity (in region of best adaptability)



days from transplanting until mature
78
91



green stage



days from transplanting until mature
115
127



red or yellow stage



days from direct seeding until mature
120
134



green stage



days from direct seeding until mature
158
170



red or yellow stage


3.
Plant



habit
semi-spreading
semi-spreading



attitude
semi-upright/
semi-upright




semi-erect (Clovis,




Sonar)



height
115 cm
135 cm



width
68 cm
60 cm



length of stem from cotyledon to first
30 cm
30 cm



flower



length of the third internode (from soil
100 mm
170 mm



surface)



length of stem
medium (Belsir,




Lamuyo)



shortened internode (in upper part)
absent (California




wonder, De




Cayenne)



for varieties without shortened
medium (Dolmi,



internodes: length of internode (on
Florian, Órias)



primary side shoots)



stem: hairiness of nodes
absent or very weak




(Arlequin)



height
medium (HRF)



basal branches
few (2-3)
(2-3)



branch flexibility
rigid (Yolo
rigid




Wonder)



stem strength (breakage resistance)
intermediate
intermediate


4.
Leaf



length of blade
medium (Atol,




Blondy, Marconi,




merit, Anthea)



width of blade
medium (Albaregia,




Balaton, Danubia,




Marconi, Merit)



width
120 mm
140 mm



length
330 mm
325 mm



petiole length
140 mm
100 mm



color
medium green
medium green



color (RHS Color Chart value)
136B
136A



intensity of green color
light (Piquant




d'Algérie,




Pusztagold)



mature leaf shape
ovate (Balico,
ovate




Sonar)



leaf and stem pubescence
absent
absent



undulation of margin
very weak
very weak



blistering
absent (Century,
very weak




Recio, Solfiane)



profile in cross section
flat (De Cayenne,




Recio)



glossiness
weak (De Cayenne,




Doux très long des




Landes)



peduncle attitude
drooping (Heldor,




Lamuyo)


5.
Flower



number of flowers per leaf axil
1
1



calyx lobes
6
6



petals
6
6



size diameter
22 mm
25 mm



corolla color
white
white



corolla throat markings

none



anther color
purple
purple



style length
same as stamen
same as stamen



self-incompatibility
absent
absent


6.
Fruit



group
Mini blocky
Bell (Yolo Wonder L.)



color (before maturity)
green (California




wonder, Lamuyo)



intensity of color (before maturity)
light



immature fruit color
medium green
medium green



immature fruit color (RHS Color Chart
139A
N134A



value)



attitude/position
drooping/pendent
drooping




(De Cayenne,




Lamuyo)



length
short (Delphin,




Petit carré doux)



diameter
narrow (Doux très




long des Landes)



ratio length/diameter
medium (Adra,




Cherry Sweet,




Daniel, Delphin,




Edino)



calyx diameter
27 mm
38 mm



fruit length
82 mm
76 mm



diameter at calyx attachment
64 mm
84 mm



diameter at mid-point
50 mm
80 mm



flesh thickness at mid-point
5 mm
7 mm



average number of fruits per plant
34
18



average fruit weight
91 gm
174 gm



shape in longitudinal section
narrowly triangular
square




(De Cayenne,




Demon)



shape in cross section (at level of
angular/triangular
circular



placenta)
(Vinedale)



sinuation of pericarp at basal part
medium (Duna,




Banán)



sinuation of pericarp excluding basal
medium (Ursus)



part



texture of surface
smooth or very
smooth




slightly wrinkled




(Milord)



color (at maturity)
orange (Ariane)



intensity of color (at maturity)
dark



mature fruit color
orange
orange



mature fruit color (RHS Color Chart
28A
28A



value)



glossiness
weak (Doux très
weak




long des Landes)



stalk cavity
absent (Corinto,




Corno di toro,




Sweet fbanana,




Sucette de




Provence)



depth of stalk cavity
shallow (Delphin,




Doux italien, Fehér,




Latino)



pedicel length
40 mm
50 mm



pedicel thickness
5 mm
15 mm



pedicel shape
curved
curved



pedicel cavity
absent
present



depth of pedicel cavity
3 mm
8 mm



stalk: length
short (Surpas, yolo




Wonder, Zenith)



stalk: thickness
thin (Sweet banana)



base shape
cupped
cupped



shape of apex
moderately
very depressed




depressed




(Quadrato a'Asti




rosso)



shape
Mini blocky
Bell (Yolo Wonder L.)



set
scattered
scattered



depth of interloculary grooves
shallow (Milord,
medium




Topgirl)



number of locules
predominantly three




(Century)



% fruits with one locule
0
0



% fruits with two locules
25
0



% fruits with three locules
50
80



% fruits with four locules
25
20



% fruits with five or more locules
0
0



thickness of flesh
medium (Fehér,




Lamuyo)



calyx: aspect
non-enveloping/
non-enveloping/




saucer-shaped
saucer-shaped




(Lamuyo, Sonar)



pungency
sweet



capsaicin in placenta
absent (Sonar)



flavor
strong pepper
mild




flavor



glossiness
moderate
dull


7.
Seed



seed cavity length
55 mm
47 mm



seed cavity diameter
40 mm
62 mm



placenta length
20 mm
25 mm



number of seeds per fruit
180
150



grams per 1000 seeds
7.9 gm



seed color
yellow
yellow


8.
Seedling



anthocyanin coloration of stem
moderate
moderate



anthocyanin coloration of nodes
moderate
moderate



stem: intensity of anthocyanin
medium (Clovis,



coloration of nodes
Lamuyo, Sonar)



plant: anthocyanin coloration of leaf
moderate
weak



plant: anthocyanin coloration of pedicel
weak
weak



plant: anthocyanin coloration of calyx
absent
absent



flower: anthocyanin coloration in
present (Lamuyo)



anther



fruit: anthocyanin coloration
absent (Lamuyo)
weak



beginning of flowering (1st flower on
early (Carré doux



2nd flowering node)
extra hâtif, Cupido,




Fehér, Flaviano,




Lito, Trophy)


9.
Resistance



to Tobamovirus Pathotype 0
present/most
present/most



(Tobacco Mosaic Virus (0))
resistant (Lamuyo,
resistant




Sonar, Yolo




Wonder)



to Tobamovirus Pathotype 1-2 (Pepper
present (Delgado,



Mosaic Virus (1-2))
Festos, Novi,




Orion)



to Tobamovirus Pathotype 1-2-3
absent/most
absent/most



(Pepper Mild Mottle Virus (1-2-3))
susceptible
susceptible




(Piperade, Yolo




Wonder)



to Curly Top Virus
most susceptible
most susceptible



to Potato Virus Y (PVY)
most resistant
most susceptible



to PVY Pathotype 0
present (Yolo Y)



to PVY Pathotype 1
absent (Yolo




Wonder, Yolo Y)



to PVY Pathotype 1-2
absent (Florida




VR2, Yolo




Wonder, Yolo Y)



to Tobacco Etch Virus
most susceptible
most susceptible



to Phytophthora capsici
absent/most
most susceptible




susceptible (Yolo




Wonder)



to Cucumber Mosiac Virus (CMV)
absent/most
most susceptible




susceptible (Yolo




Wonder)



to Pepper Spotted Wilt Virus (TSWV)
absent (Yolo




Wonder)



to Xanthomonas campestris pv.
absent/most
most susceptible



vesicatoria
susceptible




(Fehérözön, Yolo




Wonder)



to Anthracnose (Gloeosporium
most susceptible
most susceptible



piperatum)



to Cercospora Leaf Spot (Cercospors
most susceptible
most susceptible



capsici)



to Nematode (Meloidogyne incognita
most susceptible
most susceptible



acrita)



to Ripe Rot (Vermicularia capsici)
most susceptible
most susceptible



to Southern Blight (Sclerotium rolfsii)
most susceptible
most susceptible



to Verticillium Wilt (Verticillium
most susceptible
most susceptible



dahliae)





*These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention.













TABLE 2







Physiological and Morphological Characteristics of Line SMO28-1234













Comparison



CHARACTERISTIC
SMO28-1234
Variety-Boogie














1.
Species
C. annuum
C. annuum


2.
Maturity (in region of best adaptability)



days from transplanting until mature
45
50



green stage



days from transplanting until mature
70
75



red or yellow stage



days from direct seeding until mature
80
85



green stage



days from direct seeding until mature
105
110



red or yellow stage


3.
Plant



habit
compact
compact



attitude
prostrate (Delphin,
prostrate (Delphin,




Trophy)
Trophy)



height
110 cm
130 cm



width
50 cm
55 cm



length of stem from cotyledon to first
29 cm
24 cm



flower



length of the third internode (from soil
195 mm
130 mm



surface)



length of stem
short (Delphin,




Trophy)



shortened internode (in upper part)
absent (California




wonder, De




Cayenne)



for varieties without shortened
short (Bandero,



internodes: length of internode (on
Blondy, Danubia,



primary side shoots)
Tenor)



stem: hairiness of nodes
absent or very weak




(Arlequin)



height
short (Albaregia)



basal branches
few (2-3)
few (2-3)



branch flexibility
willowy (Cayenne
willowy (Cayenne




Long Red)
Long Red)



stem strength (breakage resistance)
intermediate
intermediate


4.
Leaf



length of blade
medium (Atol,




Blondy, Marconi,




Merit, Anthea)



width of blade
medium (Albaregia,




Balaton, Danubia,




Marconi, Merit)



width
135 mm
140 mm



length
215 mm
220 mm



petiole length
105 mm
95 mm



color
light green
dark green



color (RHS Color Chart value)
137A
136A



intensity of green color
light (Piquant




d'Algérie,




Pusztagold)



mature leaf shape
ovate (Balico,
broad elliptic




Sonar)
(Solario)



leaf and stem pubescence
light
light



undulation of margin
absent (De




Cayenne)



blistering
absent (Century,




Recio, Solfiane)



profile in cross section
strongly concave




(Slávy)



glossiness
very weak




(Diavolo)



peduncle attitude
drooping (Heldor,




Lamuyo)


5.
Flower



number flowers per leaf axil
1
1



calyx lobes
7
7



petals
6
7



size diameter
34
38



corolla color
white
white



anther color
purple
purple



style length
same as stamen
exceeds stamen



self-incompatibility
absent
absent


6.
Fruit



group
Mini Pepper -
Bell (Yolo Wonder L.)




Conical



color (before maturity)
green (California




wonder, Lamuyo)



intensity of color (before maturity)
medium



immature fruit color
medium green
dark green



immature fruit color (RHS Color Chart
135A
139A



value)



attitude/position
drooping/pendent
drooping/pendent




(De Cayenne,
(De Cayenne,




Lamuyo)
Lamuyo)



length
very short (Cherry




Sweet, Topgirl)



diameter
narrow (Doux très




long des Landes)



ratio length/diameter
medium (Adra,




Cherry Sweet,




Daniel, Delphin,




Edino)



calyx diameter
24 mm
32 mm



fruit length
60 mm
72 mm



diameter at calyx attachment
44 mm
80 mm



diameter at mid-point
40 mm
80 mm



flesh thickness at mid-point
6 mm
8 mm



average number of fruits per plant
20
12



% large fruits
0
90



% medium fruits
0
10



% small fruits
100
0



average fruit weight
42 g
206 g



shape in longitudinal section
moderately
cordate/heart-




triangular (Fehér,
shaped (Daniel)




Marconi)



shape in cross section (at level of
angular/triangular
angular/triangular



placenta)
(Vinedale)
(Vinedale)



sinuation of pericarp at basal part
medium (Duna,




Banán)



sinuation of pericarp excluding basal
weak (Clovis,



part
Sonar)



texture of surface
smooth or very
smooth or very




slightly wrinkled
slightly wrinkled




(Milord)
(Milord)



color (at maturity)
orange (Ariane)



intensity of color (at maturity)
dark



mature fruit color
orange
orange



mature fruit color (RHS Color Chart
28A
28A



value)



glossiness
strong (Doux
medium/moderate




italien, Trophy)
(Carré doux extra





hâtif, Lamuyo,





Sonar)



stalk cavity
present (Bingor,




Lamuyo)



depth of stalk cavity
very shallow




(Flush, Kaméleon,




Niagara)



pedicel length
40 mm
50 mm



pedicel thickness
7 mm
12 mm



pedicel shape
curved
curved



pedicel cavity
absent
present



depth of pedicel cavity
2 mm
4 mm



stalk: length
medium (Fehér,




Sonar)



stalk: thickness
thin (Sweet banana)



base shape
cupped
cupped



shape of apex
moderately
blunt




depressed




(Quadrato a'Asti




rosso)



shape
oblong (Jalapeno)
Bell (Yolo Wonder L.)



set
scattered
scattered



depth of interloculary grooves
medium (Clovis,
deep (Majister,




Lamuyo, Marconi)
Surpas)



number of locules
predominantly three




(Century)



% fruits with one locule
0
0



% fruits with two locules
2
0



% fruits with three locules
98
50



% fruits with four locules
0
45



% fruits with five or more locules
0
5



average number of locules
3
3.50



thickness of flesh
medium (Fehér,




Lamuyo)



calyx: aspect
non-enveloping/
non-enveloping/




saucer-shaped
saucer-shaped




(Lamuyo, Sonar)
(Lamuyo, Sonar)



pungency
sweet
sweet



capsaicin in placenta
absent (Sonar)



flavor
strong pepper
strong pepper flavor




flavor



glossiness
shiny
moderate


7.
Seed



seed cavity length
22 mm
45 mm



seed cavity diameter
12 mm
17 mm



placenta length
14 mm
28 mm



number of seeds per fruit
50
100



grams per 1000 seeds
7.3



seed color
yellow
yellow


8.
Seedling



anthocyanin coloration of hypocotyl
weak
moderate



anthocyanin coloration of stem
weak
moderate



anthocyanin coloration of nodes
moderate
strong (California





wonder)



stem: intensity of anthocyanin
medium (Clovis,



coloration of nodes
Lamuyo, Sonar)



plant: anthocyanin coloration of leaf
weak
weak



plant: anthocyanin coloration of pedicel
absent
weak



plant: anthocyanin coloration of calyx
absent
absent



flower: anthocyanin coloration in
present (Lamuyo)



anther



fruit: anthocyanin coloration
absent (Lamuyo)
absent (Lamuyo)



Time of maturity
very early (Koral,




Macska sárga,




Madison)


9.
Resistance



to Tobamovirus Pathotype 0
present/most
present/most



(Tobacco Mosaic Virus (0))
resistant (Lamuyo,
resistant (Lamuyo,




Sonar, Yolo
Sonar, Yolo




Wonder)
Wonder)



to Tobamovirus Pathotype 1-2 (Pepper
present (Delgado,



Mosaic Virus (1-2))
Festos, Novi,




Orion)



to Tobamovirus Pathotype 1-2-3
absent/most
absent/most



(Pepper Mild Mottle Virus (1-2-3))
susceptible
susceptible




(Piperade, Yolo
(Piperade, Yolo




Wonder)
Wonder)



to Curly Top Virus
most susceptible
most susceptible



to Potato Virus Y (PVY)
most resistant
most susceptible



to PVY Pathotype 0
present (Yolo Y)



to PVY Pathotype 1
absent (Yolo




Wonder, Yolo Y)



to PVY Pathotype 1-2
absent (Florida




VR2, Yolo




Wonder, Yolo Y)



to Tobacco Etch Virus
most susceptible
most susceptible



to Phytophthora capsici
absent/most
absent/most




susceptible (Yolo
susceptible (Yolo




Wonder)
Wonder)



to Cucumber Mosiac Virus (CMV)
absent/most
absent/most




susceptible (Yolo
susceptible (Yolo




Wonder)
Wonder)



to Pepper Spotted Wilt Virus (TSWV)
absent (Yolo




Wonder)



to Xanthomonas campestris pv.
absent/most
absent/most



vesicatoria
susceptible
susceptible




(Fehérözön, Yolo
(Fehérözön, Yolo




Wonder)
Wonder)



to Anthracnose (Gloeosporium
most susceptible
most susceptible



piperatum)



to Cercospora Leaf Spot (Cercospors
most susceptible
most susceptible



capsici)



to Nematode (Meloidogyne incognita
most susceptible
most susceptible



acrita)



to Ripe Rot (Vermicularia capsici)
most susceptible
most susceptible



to Southern Blight (Sclerotium rolfsii)
most susceptible
most susceptible



to Verticillium Wilt (Verticillium
most susceptible
most susceptible



dahliae)





*These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention.






C. Breeding Pepper Plants
One aspect of the current invention concerns methods for producing seed of pepper hybrid BS02828008 involving crossing pepper lines SMO28-1234 and SMO28-1254. Alternatively, in other embodiments of the invention, hybrid BS02828008, line SMO28-1234, or line SMO28-1254 may be crossed with itself or with any second plant. Such methods can be used for propagation of hybrid BS02828008 and/or the pepper lines SMO28-1234 and SMO28-1254, or can be used to produce plants that are derived from hybrid BS02828008 and/or the pepper lines SMO28-1234 and SMO28-1254. Plants derived from hybrid BS02828008 and/or the pepper lines SMO28-1234 and SMO28-1254 may be used, in certain embodiments, for the development of new pepper varieties.
The development of new varieties using one or more starting varieties is well known in the art. In accordance with the invention, novel varieties may be created by crossing hybrid BS02828008 followed by multiple generations of breeding according to such well known methods. New varieties may be created by crossing with any second plant. In selecting such a second plant to cross for the purpose of developing novel lines, it may be desired to choose those plants which either themselves exhibit one or more selected desirable characteristics or which exhibit the desired characteristic(s) when in hybrid combination. Once initial crosses have been made, inbreeding and selection take place to produce new varieties. For development of a uniform line, often five or more generations of selfing and selection are involved.
Uniform lines of new varieties may also be developed by way of double-haploids. This technique allows the creation of true breeding lines without the need for multiple generations of selfing and selection. In this manner true breeding lines can be produced in as little as one generation. Haploid embryos may be produced from microspores, pollen, anther cultures, or ovary cultures. The haploid embryos may then be doubled autonomously, or by chemical treatments (e.g. colchicine treatment). Alternatively, haploid embryos may be grown into haploid plants and treated to induce chromosome doubling. In either case, fertile homozygous plants are obtained. In accordance with the invention, any of such techniques may be used in connection with a plant of the invention and progeny thereof to achieve a homozygous line.
Backcrossing can also be used to improve an inbred plant. Backcrossing transfers a specific desirable trait from one inbred or non-inbred source to an inbred that lacks that trait. This can be accomplished, for example, by first crossing a superior inbred (A) (recurrent parent) to a donor inbred (non-recurrent parent), which carries the appropriate locus or loci for the trait in question. The progeny of this cross are then mated back to the superior recurrent parent (A) followed by selection in the resultant progeny for the desired trait to be transferred from the non-recurrent parent. After five or more backcross generations with selection for the desired trait, the progeny have the characteristic being transferred, but are like the superior parent for most or almost all other loci. The last backcross generation would be selfed to give pure breeding progeny for the trait being transferred.
The plants of the present invention are particularly well suited for the development of new lines based on the elite nature of the genetic background of the plants. In selecting a second plant to cross with BS02828008 and/or pepper lines SMO28-1234 and SMO28-1254 for the purpose of developing novel pepper lines, it will typically be preferred to choose those plants which either themselves exhibit one or more selected desirable characteristics or which exhibit the desired characteristic(s) when in hybrid combination. Examples of desirable traits may include, in specific embodiments, high seed yield, high seed germination, seedling vigor, high fruit yield, disease tolerance or resistance, and adaptability for soil and climate conditions. Consumer-driven traits, such as a fruit shape, color, texture, and taste are other examples of traits that may be incorporated into new lines of pepper plants developed by this invention.
D. Performance Characteristics
As described above, hybrid BS02828008 exhibits desirable agronomic traits. The performance characteristics of BS02828008, SMO28-1234 and SMO28-1254 were the subject of an objective analysis of the performance traits relative to other varieties. The results of the analysis are presented below.







TABLE 3







Performance Characteristics For BS02828008























Fruit size length






Tobamo virus
Tobamo virus
Potato Virus Y
Antho-
Fruit color at
(cm) × width
Longitundinal


Hybrid
Source
P0 resistance
P1.2 resistance
P0 resistance
cyaninless
maturity
(cm)
section shape
Locules





BS02828008
Seminis
yes
yes
yes
no
dark
8.2 × 5.0
narrowly
3








orange

triangular


E 499524
Enza
yes
no
yes
no
light red
5 × 2
trapezoidal
2 or 3



Laden


E 499526
Enza
yes
no
yes
no
light
4 × 2
trapezoidal
2 or 3



Laden




yellow


E 499531
Enza
yes
no
yes
no
medium
4 × 2
trapezoidal
2 or 3



Laden




orange


Ferrari
Enza
yes


no
dark red
7.6 × 3.6
rectangular
2 or 3



Laden


Derby
DeRuiter


no
no
orange-
8.8 × 7.0
square
3








yellow


Boogie
Rijk
yes

no
no
orange
7.6 × 8.0
square
3



Zwaan


ABSA
Seminis
yes
no
yes
no
orange
9.4 × 4.0
narrowly
2


Orange







triangular
















TABLE 4







Performance Characteristics For SMO28-1234












Tobamo virus






P0.1.2
Potato Virus Y
Fruit
Fruit



resistance
P0 resistance
diameter
shape















SMO28-1234
Resistant
Resistant
4.0 cm
Oblong


Boogie
Susceptible
Susceptible
8.0 cm
Blocky
















TABLE 5







Performance Characteristics For SMO28-1254










Tobamo virus P0.1.2




resistance
Potato Virus Y P0 resistance













SMO28-1254
Resistant
Resistant


Boogie
Susceptible
Susceptible









E. Further Embodiments of the Invention
In certain aspects of the invention, plants described herein are provided modified to include at least a first desired heritable trait. Such plants may, in one embodiment, be developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a variety are recovered in addition to a genetic locus transferred into the plant via the backcrossing technique. The term single locus converted plant as used herein refers to those pepper plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a variety are recovered in addition to the single locus transferred into the variety via the backcrossing technique. By essentially all of the morphological and physiological characteristics, it is meant that the characteristics of a plant are recovered that are otherwise present when compared in the same environment, other than an occasional variant trait that might arise during backcrossing or direct introduction of a transgene.
Backcrossing methods can be used with the present invention to improve or introduce a characteristic into the present variety. The parental pepper plant which contributes the locus for the desired characteristic is termed the nonrecurrent or donor parent. This terminology refers to the fact that the nonrecurrent parent is used one time in the backcross protocol and therefore does not recur. The parental pepper plant to which the locus or loci from the nonrecurrent parent are transferred is known as the recurrent parent as it is used for several rounds in the backcrossing protocol.
In a typical backcross protocol, the original variety of interest (recurrent parent) is crossed to a second variety (nonrecurrent parent) that carries the single locus of interest to be transferred. The resulting progeny from this cross are then crossed again to the recurrent parent and the process is repeated until a pepper plant is obtained wherein essentially all of the morphological and physiological characteristics of the recurrent parent are recovered in the converted plant, in addition to the single transferred locus from the nonrecurrent parent.
The selection of a suitable recurrent parent is an important step for a successful backcrossing procedure. The goal of a backcross protocol is to alter or substitute a single trait or characteristic in the original variety. To accomplish this, a single locus of the recurrent variety is modified or substituted with the desired locus from the nonrecurrent parent, while retaining essentially all of the rest of the desired genetic, and therefore the desired physiological and morphological constitution of the original variety. The choice of the particular nonrecurrent parent will depend on the purpose of the backcross; one of the major purposes is to add some commercially desirable trait to the plant. The exact backcrossing protocol will depend on the characteristic or trait being altered and the genetic distance between the recurrent and nonrecurrent parents. Although backcrossing methods are simplified when the characteristic being transferred is a dominant allele, a recessive allele, or an additive allele (between recessive and dominant), may also be transferred. In this instance it may be necessary to introduce a test of the progeny to determine if the desired characteristic has been successfully transferred.
In one embodiment, progeny pepper plants of a backcross in which a plant described herein is the recurrent parent comprise (i) the desired trait from the non-recurrent parent and (ii) all of the physiological and morphological characteristics of pepper the recurrent parent as determined at the 5% significance level when grown in the same environmental conditions.
New varieties can also be developed from more than two parents. The technique, known as modified backcrossing, uses different recurrent parents during the backcrossing. Modified backcrossing may be used to replace the original recurrent parent with a variety having certain more desirable characteristics or multiple parents may be used to obtain different desirable characteristics from each.
Many single locus traits have been identified that are not regularly selected for in the development of a new inbred but that can be improved by backcrossing techniques. Single locus traits may or may not be transgenic; examples of these traits include, but are not limited to, herbicide resistance, resistance to bacterial, fungal, or viral disease, insect resistance, modified fatty acid or carbohydrate metabolism, and altered nutritional quality. These comprise genes generally inherited through the nucleus.
Direct selection may be applied where the single locus acts as a dominant trait. For this selection process, the progeny of the initial cross are assayed for viral resistance and/or the presence of the corresponding gene prior to the backcrossing. Selection eliminates any plants that do not have the desired gene and resistance trait, and only those plants that have the trait are used in the subsequent backcross. This process is then repeated for all additional backcross generations.
Selection of pepper plants for breeding is not necessarily dependent on the phenotype of a plant and instead can be based on genetic investigations. For example, one can utilize a suitable genetic marker which is closely genetically linked to a trait of interest. One of these markers can be used to identify the presence or absence of a trait in the offspring of a particular cross, and can be used in selection of progeny for continued breeding. This technique is commonly referred to as marker assisted selection. Any other type of genetic marker or other assay which is able to identify the relative presence or absence of a trait of interest in a plant can also be useful for breeding purposes. Procedures for marker assisted selection are well known in the art. Such methods will be of particular utility in the case of recessive traits and variable phenotypes, or where conventional assays may be more expensive, time consuming or otherwise disadvantageous. Types of genetic markers which could be used in accordance with the invention include, but are not necessarily limited to, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., 1990), Randomly Amplified Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang et al., 1998).
F. Plants Derived by Genetic Engineering
Many useful traits that can be introduced by backcrossing, as well as directly into a plant, are those which are introduced by genetic transformation techniques. Genetic transformation may therefore be used to insert a selected transgene into a plant of the invention or may, alternatively, be used for the preparation of transgenes which can be introduced by backcrossing. Methods for the transformation of plants that are well known to those of skill in the art and applicable to many crop species include, but are not limited to, electroporation, microprojectile bombardment, Agrobacterium-mediated transformation and direct DNA uptake by protoplasts.
To effect transformation by electroporation, one may employ either friable tissues, such as a suspension culture of cells or embryogenic callus or alternatively one may transform immature embryos or other organized tissue directly. In this technique, one would partially degrade the cell walls of the chosen cells by exposing them to pectin-degrading enzymes (pectolyases) or mechanically wound tissues in a controlled manner.
An efficient method for delivering transforming DNA segments to plant cells is microprojectile bombardment. In this method, particles are coated with nucleic acids and delivered into cells by a propelling force. Exemplary particles include those comprised of tungsten, platinum, and preferably, gold. For the bombardment, cells in suspension are concentrated on filters or solid culture medium. Alternatively, immature embryos or other target cells may be arranged on solid culture medium. The cells to be bombarded are positioned at an appropriate distance below the macroprojectile stopping plate.
An illustrative embodiment of a method for delivering DNA into plant cells by acceleration is the Biolistics Particle Delivery System, which can be used to propel particles coated with DNA or cells through a screen, such as a stainless steel or Nytex screen, onto a surface covered with target cells. The screen disperses the particles so that they are not delivered to the recipient cells in large aggregates. Microprojectile bombardment techniques are widely applicable, and may be used to transform virtually any plant species.
Agrobacterium-mediated transfer is another widely applicable system for introducing gene loci into plant cells. An advantage of the technique is that DNA can be introduced into whole plant tissues, thereby bypassing the need for regeneration of an intact plant from a protoplast. Modern Agrobacterium transformation vectors are capable of replication in E. coli as well as Agrobacterium, allowing for convenient manipulations (Klee et al., 1985). Moreover, recent technological advances in vectors for Agrobacterium-mediated gene transfer have improved the arrangement of genes and restriction sites in the vectors to facilitate the construction of vectors capable of expressing various polypeptide coding genes. The vectors described have convenient multi-linker regions flanked by a promoter and a polyadenylation site for direct expression of inserted polypeptide coding genes. Additionally, Agrobacterium containing both armed and disarmed Ti genes can be used for transformation.
In those plant strains where Agrobacterium-mediated transformation is efficient, it is the method of choice because of the facile and defined nature of the gene locus transfer. The use of Agrobacterium-mediated plant integrating vectors to introduce DNA into plant cells is well known in the art (Fraley et al., 1985; U.S. Pat. No. 5,563,055).
Transformation of plant protoplasts also can be achieved using methods based on calcium phosphate precipitation, polyethylene glycol treatment, electroporation, and combinations of these treatments (see, e.g., Potrykus et al., 1985; Omirulleh et al., 1993; Fromm et al., 1986; Uchimiya et al., 1986; Marcotte et al., 1988). Transformation of plants and expression of foreign genetic elements is exemplified in Choi et al. (1994), and Ellul et al. (2003).
A number of promoters have utility for plant gene expression for any gene of interest including but not limited to selectable markers, scoreable markers, genes for pest tolerance, disease resistance, nutritional enhancements and any other gene of agronomic interest. Examples of constitutive promoters useful for plant gene expression include, but are not limited to, the cauliflower mosaic virus (CaMV) P-35S promoter, which confers constitutive, high-level expression in most plant tissues (see, e.g., Odel et al., 1985), including in monocots (see, e.g., Dekeyser et al., 1990; Terada and Shimamoto, 1990); a tandemly duplicated version of the CaMV 35S promoter, the enhanced 35S promoter (P-e35S); 1 the nopaline synthase promoter (An et al., 1988); the octopine synthase promoter (Fromm et al., 1989); and the figwort mosaic virus (P-FMV) promoter as described in U.S. Pat. No. 5,378,619 and an enhanced version of the FMV promoter (P-eFMV) where the promoter sequence of P-FMV is duplicated in tandem; the cauliflower mosaic virus 19S promoter; a sugarcane bacilliform virus promoter; a commelina yellow mottle virus promoter; and other plant DNA virus promoters known to express in plant cells.
A variety of plant gene promoters that are regulated in response to environmental, hormonal, chemical, and/or developmental signals can also be used for expression of an operably linked gene in plant cells, including promoters regulated by (1) heat (Callis et al., 1988), (2) light (e.g., pea rbcS-3A promoter, Kuhlemeier et al., 1989; maize rbcS promoter, Schaffner and Sheen, 1991; or chlorophyll a/b-binding protein promoter, Simpson et al., 1985), (3) hormones, such as abscisic acid (Marcotte et al., 1989), (4) wounding (e.g., wunl, Siebertz et al., 1989); or (5) chemicals such as methyl jasmonate, salicylic acid, or Safener. It may also be advantageous to employ organ-specific promoters (e.g., Roshal et al., 1987; Schernthaner et al., 1988; Bustos et al., 1989).
Exemplary nucleic acids which may be introduced to plants of this invention include, for example, DNA sequences or genes from another species, or even genes or sequences which originate with or are present in the same species, but are incorporated into recipient cells by genetic engineering methods rather than classical reproduction or breeding techniques. However, the term “exogenous” is also intended to refer to genes that are not normally present in the cell being transformed, or perhaps simply not present in the form, structure, etc., as found in the transforming DNA segment or gene, or genes which are normally present and that one desires to express in a manner that differs from the natural expression pattern, e.g., to over-express. Thus, the term “exogenous” gene or DNA is intended to refer to any gene or DNA segment that is introduced into a recipient cell, regardless of whether a similar gene may already be present in such a cell. The type of DNA included in the exogenous DNA can include DNA which is already present in the plant cell, DNA from another plant, DNA from a different organism, or a DNA generated externally, such as a DNA sequence containing an antisense message of a gene, or a DNA sequence encoding a synthetic or modified version of a gene.
Many hundreds if not thousands of different genes are known and could potentially be introduced into a pepper plant according to the invention. Non-limiting examples of particular genes and corresponding phenotypes one may choose to introduce into a pepper plant include one or more genes for insect tolerance, such as a Bacillus thuringiensis (B.t.) gene, pest tolerance such as genes for fungal disease control, herbicide tolerance such as genes conferring glyphosate tolerance, and genes for quality improvements such as yield, nutritional enhancements, environmental or stress tolerances, or any desirable changes in plant physiology, growth, development, morphology or plant product(s). For example, structural genes would include any gene that confers insect tolerance including but not limited to a Bacillus insect control protein gene as described in WO 99/31248, herein incorporated by reference in its entirety, U.S. Pat. No. 5,689,052, herein incorporated by reference in its entirety, U.S. Pat. Nos. 5,500,365 and 5,880,275, herein incorporated by reference in their entirety. In another embodiment, the structural gene can confer tolerance to the herbicide glyphosate as conferred by genes including, but not limited to Agrobacterium strain CP4 glyphosate resistant EPSPS gene (aroA:CP4) as described in U.S. Pat. No. 5,633,435, herein incorporated by reference in its entirety, or glyphosate oxidoreductase gene (GOX) as described in U.S. Pat. No. 5,463,175, herein incorporated by reference in its entirety.
Alternatively, the DNA coding sequences can affect these phenotypes by encoding a non-translatable RNA molecule that causes the targeted inhibition of expression of an endogenous gene, for example via antisense- or cosuppression-mediated mechanisms (see, for example, Bird et al., 1991). The RNA could also be a catalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desired endogenous mRNA product (see for example, Gibson and Shillito, 1997). Thus, any gene which produces a protein or mRNA which expresses a phenotype or morphology change of interest is useful for the practice of the present invention.
G. Definitions
In the description and tables herein, a number of terms are used. In order to provide a clear and consistent understanding of the specification and claims, the following definitions are provided:
Allele: Any of one or more alternative forms of a gene locus, all of which alleles relate to one trait or characteristic. In a diploid cell or organism, the two alleles of a given gene occupy corresponding loci on a pair of homologous chromosomes.
Backcrossing: A process in which a breeder repeatedly crosses hybrid progeny, for example a first generation hybrid (F1), back to one of the parents of the hybrid progeny. Backcrossing can be used to introduce one or more single locus conversions from one genetic background into another.
Crossing: The mating of two parent plants.
Cross-pollination: Fertilization by the union of two gametes from different plants.
Diploid: A cell or organism having two sets of chromosomes.
Emasculate: The removal of plant male sex organs or the inactivation of the organs with a cytoplasmic or nuclear genetic factor or a chemical agent conferring male sterility.
Enzymes: Molecules which can act as catalysts in biological reactions.
F1 Hybrid: The first generation progeny of the cross of two nonisogenic plants.
Genotype: The genetic constitution of a cell or organism.
Haploid: A cell or organism having one set of the two sets of chromosomes in a diploid.
Linkage: A phenomenon wherein alleles on the same chromosome tend to segregate together more often than expected by chance if their transmission was independent.
Marker: A readily detectable phenotype, preferably inherited in codominant fashion (both alleles at a locus in a diploid heterozygote are readily detectable), with no environmental variance component, i.e., heritability of 1.
Phenotype: The detectable characteristics of a cell or organism, which characteristics are the manifestation of gene expression.
Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer to genetic loci that control to some degree numerically representable traits that are usually continuously distributed.
Resistance: As used herein, the terms “resistance” and “tolerance” are used interchangeably to describe plants that show no symptoms to a specified biotic pest, pathogen, abiotic influence or environmental condition. These terms are also used to describe plants showing some symptoms but that are still able to produce marketable product with an acceptable yield. Some plants that are referred to as resistant or tolerant are only so in the sense that they may still produce a crop, even though the plants are stunted and the yield is reduced.
Regeneration: The development of a plant from tissue culture.
Royal Horticultural Society (RHS) color chart value: The RHS color chart is a standardized reference which allows accurate identification of any color. A color's designation on the chart describes its hue, brightness and saturation. A color is precisely named by the RHS color chart by identifying the group name, sheet number and letter, e.g., Yellow-Orange Group 19A or Red Group 41B.
Self-pollination: The transfer of pollen from the anther to the stigma of the same plant.
Single Locus Converted (Conversion) Plant: Plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a pepper variety are recovered in addition to the characteristics of the single locus transferred into the variety via the backcrossing technique and/or by genetic transformation.
Substantially Equivalent: A characteristic that, when compared, does not show a statistically significant difference (e.g., p=0.05) from the mean.
Tissue Culture: A composition comprising isolated cells of the same or a different type or a collection of such cells organized into parts of a plant.
Transgene: A genetic locus comprising a sequence which has been introduced into the genome of a pepper plant by transformation.
H. Deposit Information
A deposit of pepper hybrid BS02828008 and inbred parent lines SMO28-1234 and SMO28-1254, disclosed above and recited in the claims, has been made with the American Type Culture Collection (ATCC), 10801 University Blvd., Manassas, Va. 20110-2209. The date of deposit was May 26, 2009. The accession numbers for those deposited seeds of pepper hybrid BS02828008 and inbred parent lines SMO28-1234 and SMO28-1254 are ATCC Accession Number PTA-10069, ATCC Accession Number PTA-10074, and ATCC Accession Number PTA-10075, respectively. Upon issuance of a patent, all restrictions upon the deposits will be removed, and the deposits are intended to meet all of the requirements of 37 C.F.R. §1.801-1.809. The deposits will be maintained in the depository for a period of 30 years, or 5 years after the last request, or for the effective life of the patent, whichever is longer, and will be replaced if necessary during that period.
Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity and understanding, it will be obvious that certain changes and modifications may be practiced within the scope of the invention, as limited only by the scope of the appended claims.
All references cited herein are hereby expressly incorporated herein by reference.
REFERENCES
The following references, to the extent that they provide exemplary procedural or other details supplementary to those set forth herein, are specifically incorporated herein by reference:

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## Claims

1. A pepper plant comprising at least a first set of the chromosomes of pepper line SMO28-1254, a sample of seed of said line having been deposited under ATCC Accession Number PTA-10075.

2. A pepper seed comprising at least a first set of the chromosomes of pepper line SMO28-1254, a sample of seed of said line having been deposited under ATCC Accession Number PTA-10075.

3. The pepper plant of claim 1, which is an inbred.

4. The pepper plant of claim 1, which is a hybrid.

5. The pepper plant of claim 4, wherein the hybrid plant is pepper hybrid BS02828008, a sample of seed of said hybrid BS02828008 having been deposited under ATCC Accession Number PTA-10069.

6. The plant of claim 1, wherein the plant is a plant of pepper line SMO28-1254.

7. A plant part of the pepper plant of claim 1.

8. The plant part of claim 7, further defined as a leaf, a ovule, pollen, a fruit, or a cell.

9. A pepper plant having all the physiological and morphological characteristics of the pepper plant of claim 5.

10. A pepper plant having all the physiological and morphological characteristics of the pepper plant of claim 6.

11. A tissue culture of regenerable cells of the pepper plant of claim 1.

12. The tissue culture according to claim 11, comprising cells or protoplasts from a plant part selected from the group consisting of embryos, meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistil, flower, seed and stalks.

13. A pepper plant regenerated from the tissue culture of claim 12, wherein said plant has all the physiological and morphological characteristics of the pepper plant comprising at least a first set of the chromosomes of pepper line SMO28-1254, a sample of seed of said line having been deposited under ATCC Accession Number PTA-10075.

14. A method of vegetatively propagating the pepper plant of claim 1 comprising the steps of:
(a) collecting tissue capable of being propagated from a pepper plant according to claim 1;
(b) cultivating said tissue to obtain proliferated shoots; and
(c) rooting said proliferated shoots to obtain rooted plantlets.

15. The method of claim 14, further comprising growing at least a first pepper plant from said rooted plantlets.

16. A method of introducing a desired trait into a pepper line, comprising:
(a) utilizing as a recurrent parent a plant of pepper line SMO28-1254, by crossing a plant of pepper line SMO28-1254 with a second donor pepper plant that comprises a desired trait to produce F1 progeny, a sample of seed of said line having been deposited under ATCC Accession Number PTA-10075;
(b) selecting an F1 progeny that comprises the desired trait;
(c) backcrossing the selected F1 progeny with a plant of the same pepper line used as the recurrent parent in step (a), to produce backcross progeny;
(d) selecting backcross progeny comprising the desired trait and the physiological and morphological characteristics of the recurrent parent pepper line used in step (a); and
(e) repeating steps (c) and (d) three or more times to produce selected fourth or higher backcross progeny that comprise the desired trait, and otherwise comprise essentially all of the morphological and physiological characteristics of the recurrent parent pepper line used in step (a).

17. A pepper plant produced by the method of claim 16.

18. A method of producing a pepper plant comprising an added trait, the method comprising introducing a transgene conferring the trait into a plant of pepper hybrid BS02828008 or pepper line SMO28-1254, a sample of seed of said hybrid and line having been deposited under ATCC Accession Number PTA-10069 and ATCC Accession Number PTA-10075 respectively.

19. A pepper plant produced by the method of claim 18.

20. A method of determining the genotype of the plant of claim 1 comprising obtaining a sample of nucleic acids from said plant and detecting in said nucleic acids a plurality of polymorphisms.

21. The method of claim 20, further comprising the step of storing the results of detecting the plurality of polymorphisms on a computer readable medium.

22. The plant of claim 1, further comprising a transgene.

23. The plant of claim 22, wherein the transgene confers a trait selected from the group consisting of male sterility, herbicide tolerance, insect resistance, pest resistance, disease resistance, modified fatty acid metabolism, environmental stress tolerance, modified carbohydrate metabolism and modified protein metabolism.

24. The plant of claim 1, further comprising a single locus conversion, wherein the plant otherwise comprises essentially all of the morphological and physiological characteristics of the plant comprising at least a first set of the chromosomes of pepper line SMO28-1254, a sample of seed of said line having been deposited under ATCC Accession Number PTA-10075.

25. The plant of claim 24, wherein the single locus conversion confers a trait selected from the group consisting of male sterility, herbicide tolerance, insect resistance, pest resistance, disease resistance, modified fatty acid metabolism, environmental stress tolerance, modified carbohydrate metabolism and modified protein metabolism.

26. A method for producing a seed of a pepper plant derived from at least one of pepper hybrid BS02828008 or pepper line SMO28-1254 comprising the steps of:
(a) crossing a pepper plant of hybrid BS02828008 or line SMO28-1254 with itself or a second pepper plant; a sample of seed of said hybrid and line having been deposited under ATCC Accession Number PTA-10069 and ATCC Accession Number PTA-10075, respectively; and
(b) allowing seed of a hybrid BS02828008 or line SMO28-1254-derived pepper plant to form.

27. The method of claim 26, further comprising the steps of:
(c) selfing a pepper plant grown from said hybrid BS02828008 or line SMO28-1254-derived pepper seed to yield additional hybrid BS02828008 or line SMO28-1254-derived pepper seed;
(d) growing said additional hybrid BS02828008 or line SMO28-1254-derived pepper seed of step (c) to yield additional hybrid BS02828008 or line SMO28-1254-derived pepper plants; and
(e) repeating the crossing and growing steps of (c) and (d) to generate at least a first further hybrid BS02828008 or line SMO28-1254-derived pepper plant.

28. The method of claim 26, wherein the second pepper plant is of an inbred pepper line.

29. The method of claim 26, comprising crossing line SMO28-1234 with line SMO28-1254, a sample of seed of said lines having been deposited under ATCC Accession Number PTA-10074, and ATCC Accession Number PTA-10075, respectively.

30. The method of claim 27, further comprising:
(f) crossing the further hybrid BS02828008 or line SMO28-1254-derived pepper plant with a second pepper plant to produce seed of a hybrid progeny plant.

31. A hybrid pepper seed produced by the method of claim 29.

32. A pepper plant produced by growing the seed of claim 31.

33. A plant part of the pepper plant of claim 32.

34. The plant part of claim 33, further defined as a leaf, a flower, a fruit, an ovule, pollen, or a cell.

35. A method of producing a pepper seed comprising crossing the plant of claim 1 with itself or a second pepper plant and allowing seed to form.

36. A method of producing a pepper fruit comprising:
(a) obtaining a plant according to claim 1, wherein the plant has been cultivated to maturity; and
(b) collecting a pepper from the plant.